CD11b is implicated in various adhesive interactions of monocytes, macrophages and granulocytes as well as in mediating the uptake of complement coated particles. It is identical to CR3, the receptor for the iC3b fragment of the third complement component. It probably recognizes the RGD peptide in C3b. CD11b is also a receptor for fibrinogen, factor X, and ICAM1. It recognizes P1 and P2 peptides of fibrinogen gamma chain. The Mac1 CD11b antigen is present on macrophages, granulocytes, natural killer cells, and blood monocytes. CD11b is expressed on 8% of spleen cells, 44% of bone marrow cells, and less than 1% of thymocytes, and is commonly used as a microglial marker in nervous tissue.Description: The CBRM1/5 monoclonal antibody reacts with an activation-specific epitope of human Mac-1. CBRM1/5 binds a subset of Mac-1 molecules on neutrophils and monocytes after stimulation with chemoattractants or phorbol esters but does not recognize Mac-1 on resting myeloid cells. Through interactions with its ligands, Mac-1 participates in adhesive cell interactions. The epitope recognized by this mAb localizes to the I domain on the \alpha chain of Mac-1 very close to the ligand binding site in a region that is widely exposed. CBRM1/5 blocks Mac-1 dependent adhesion to fibrinogen and ICAM-1 and inhibits chemoattractant-stimulated adhesion of eosinophils to the Intercellular Adhesion Molecule-1 (ICAM-1). It should be noted that low level activation may occur during processing of freshly drawn blood. Therefore the CBRM1.5 antibody may exhibit some binding to Mac-1 in these unstimulated samples. However, higher levels of Mac-1 expression are observed in activated samples when compared to unstimulated cells. Applications Reported: This CBRM1/5 antibody has been reported for use in flow cytometric analysis. Applications Tested: This CBRM1/5 antibody has been pre-titrated and tested by flow cytometric analysis of resting and activated normal human peripheral blood cells. This can be used at 5µL (0.125µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100µL cell sample + 100µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.Filtration: 0.2µm post-manufacturing filtered.
|CD11b (activation epitope)|
|Store at 2-8°C. Do not freeze. Light-sensitive material.|
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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